Can you dilute primers in water?

Can you dilute primers in water?

Well, DEPC water can be acidic which may cause the oligo to degrade. TE buffer (10mM Tris: 0.1mM EDTA; pH 8.0) is the safest to dilute primers. After adding water or buffer, briefly vortex the tube, but do not centrifuge it.

What should I resuspend primers in?

We recommend resuspending oligos in a TE buffer solution, such as IDTE, to maintain a constant pH that supports oligo stability (IDTE is available from IDT at pH 7.5 and pH 8.0). Alternatively, resuspend oligos in nuclease-free, sterile water, pH 7.0 (HPLC-grade is preferable; available from IDT).

Do I dilute primers in water or TE?

Either TE buffer or nuclease-free water can be used for primer dilution. If primers are to be stored long-term, however, TE buffer may be the better choice since it can help prevent degradation.

How do you dissolve lyophilized primers?

To make a typical 100 microMolar (100X) stock concentration of primers, dissolve the primers in a volume of sterile distilled water that is 10X the amount of nmoles in the tube, using microliters of water. This value is printed on the side of the tube.

How do you dissolve primers?

You can dissolve primers in: (1) Autoclaved milli-Q grade water: Add worm water (65 temp), and incubate at 65 temp in dry/water bath for ~10 min with the occasional mixing by vortex. (2) 10 mM of TE (pH-8) by vortex and proper mixing, put tubes overnight at 4 temp will enhance primer dissolution.

How do you dilute primers?

Simply 10X water (ul) of your primer (nmol) to be added to make 100uM. then the diluted concentration of primer will be 100 uM. Enjoy learning.

Can you use water instead of TE buffer?

If you “just” have samples from cells etc. that are easily reproducible or you don’t need them for longer time, you can just use water! The pH of TE buffer is slightly basic however water have slightly acidic pH.

How do you resuspend a dry primer?

This can usually be found on the tube itself or the primer sheet supplied with the order. For every 1 nmoles, add 10 μL of PCR-grade water. For example, if a primer states 19.4 nmoles, then add 194 μL of PCR-grade water. Mix the solution by vortexing to reconstitute the primers.

Why do we resuspend primer stocks in TE buffer rather than water?

TE is a good choice to resuspend high-concentration stock DNA (like 100uM PCR primers) because you know A) it will “protect” your DNA long-term by buffering and chelation, and B) you can dilute high concentration TE stocks to working concentrations with H2O later, simultaneously diluting TE/EDTA concentration.

How long are lyophilized primers good for?

Lyophilised primers when stored at -20ºC are often good for up to 18 months, but this may vary depending on supplier. In lypholized stored under perfectly dry conditions, primers can be safely stored at RT or better at -20C for extended period may be upto 5 years.

How do you reconstitute primers Sigma?

Follow these steps:

1. Resuspend the oligonucleotide in 400 µL of water or buffer.
2. Dilute 12 µL into 988 µL of sterile, nuclease-free water.
3. Take an A260 reading of the 1 mL sample in a cuvette.
4. Ensure the OD value is in the linear range (~0.1 to 1 OD).
5. Multiply the OD of the sample volume by the dilution factor.

Which is the best buffer For resuspension of primers?

The proper choice of buffer will depend on the intended application of the primers, some common ones are: Invitrogen recommends the following reconstitution procedure – Centrifuge the tube for a few seconds to get all the DNA to the bottom of the tube. Resuspend in TE buffer, pH 8.0 at a concentration greater than 10μM.

Can You resuspend vortex primer in TE buffer?

Resuspend in TE buffer, pH 8.0 at a concentration greater than 10μM. Allow to sit for 2mins, then vortex for 15s. However, if you are using primers for PCR /Sequencing you may want to resuspend in water or 100nM Tris, since the EDTA in TE buffer chelates Mg 2+ ions inhibiting PCR.

How to resuspend my IDT primers, step by step?

How do I resuspend my IDT primers? 1 Find the oligo yield information (in nmol) on the tube label or specification sheet. 2 Multiply this number by 10. 3 The resulting product is the amount of buffer needed, in µL, to prepare a 100 µM solution.

What kind of water do you use to dilute primers?

Use PCR-grade water (DNase- and RNase-free) to reconstitute and dilute your primers. To make a 10 μM working primer solution, follow these steps: Add 10 μL of primer stock solution to an RNase- and DNAse-free tube. Add 90 μL of PCR-grade water.